Purification, kinetic parameters, and isoforms of polyphenol oxidase from “Xushu 22” sweet potato skin

Abstract

We purified and compared the polyphenol oxidase (PPO) isoenzymes present in "Xushu 22," a sweet potato. A membrane-bound form (mPPO) and two soluble forms (sPPO1 and sPPO2) were identified and purified using ammonium sulphate precipitation, ion exchange chromatography, gel filtration chromatography, and mass spectrometer. The three PPO isoforms were characterized enzymatically. The specific activity of mPPO was significantly higher than that of the two sPPO isoforms, being 24.55- and 13.89-fold higher than sPPO1 and sPPO2. The preferred substrates for mPPO and the two sPPOs were catechol and chlorogenic acid, respectively. They can be efficiently and safely applied to phenolic wastewater treatment after being immobilized. Both mPPO and the two sPPOs were rapidly inactivated under acid or base conditions and were unstable at 65°C. The most effective inhibitors of mPPO, sPPO1, and sPPO2 were glutathione, ascorbic acid, and L-cysteine, respectively. PRACTICAL APPLICATIONS: According to incomplete statistics, about 5% of sweet potatoes in china are wasted due to enzymatic browning every year. PPO was regarded as a key molecule contributing to enzymatic browning in fruits and vegetables during ripening, processing, and storage and responsible for economic and industrial loss. It's great importance to characterize the PPO from "Xushu 22" sweet potato and provide data on its inactivation. The three PPO isoforms were purified and identified by chromatography and mass spectrometer. This study will provide useful informations to have a better command of PPO from "Xushu 22" sweet potato and give ideals to solve the browning of sweet potato.